Acta Scientific NUTRITIONAL HEALTH (ISSN:2582-1423) Volume 5 Issue 12 December 2021 Effect of pH and Temperature on the Isolation, Purification and Characterization of α-Amylase from Aspergillus niger Produced from Pigeon Pea Adegbanke OR1*, Adepomola AF1, Adeniran EA1 and Bamidele OS2 1 Department of Food Science and Technology, Federal University of Technology, Akure, Ondo State, Nigeria 2 Department of Biochemistry, Federal University of Technology, Akure, Ondo State, Nigeria *Corresponding Author: Adegbanke OR, Department of Food Science and Technol- ogy, Federal University of Technology, Akure, Ondo State, Nigeria. Research Article Received: September 22, 2021 Published: © All rights are reserved by Adegbanke OR., et al. Abstract α-Amylase an industrially used enzyme can be obtained from Aspergillus niger and can be produced from food sources such as pigeon pea. α-Amylase was produced from Aspergillus niger isolated from pigeon pea, purified and characterized. This process was achieved using ammonium sulphate, ion exchange DEAE column and gel filtration (Sephadex A-50 and sephadex G-100) chroma- tography. The effect of temperature and pH was determined. Ammonium sulphate precipitation results showed that the highest specific α-amylase activity was (1.01 U/mL. mg) obtained at 11.27% saturation level, with a purity of 1.81-fold of the crude extract and yielding 1.00%. Further purification using gel filtration increased the enzyme purity and yielding 8.94-fold relative to the crude extract 3.01% and yielding Specific activity after purification was 4.99 U/mg. Having varied temperature from 30-80 o C, the optimum temperature was obtained at 50°C. The pH ranged from 3-10, at a concentration of 0.01 M which was prepared using different buffer systems, the enzyme was found stable at pH of 6. Therefore α -amylase produced from Aspergillus niger can be exploited for potential usage for industrial applications of enzymes in a wide range of production and its application in food processing. Keywords: α-Amylase; Aspergillus niger; Pigeon Pea; Purification; Characterization; pH; Temperature Introduction The demand for industrial enzymes has been steadily increas- ing. This is primarily due to an increase in the demand for long- term solutions. Microbial sources have been the most important and useful sources of many of these enzymes [1] and the use of cheap and underutilized crops that is drought resistant such as pigeon pea has a huge advantage in reducing enzyme production cost. Aspergillus niger is a common fungus in nature that belongs to the Aspergillus genus. It is an important industrial fermentation microorganism that is widely used for the production of industrial enzymes and organic acids [2]. It is usually found as saprophytes and are most commonly found on stored grain, dead leaves, and other decaying vegetation. It appears in the form of very dark brown patches or carbon black and a very important group of mi- croorganisms that dominate solid state fermentation. It possesses some very special properties such as good tolerance to less avail- ability of water and capacity to spread over and into the solid sub- strate, therefore, it is extensively grown and used in food industry for the production of a variety of enzymes such as α-amylases, amyloglucosidases, cellulases, lactase and acid proteases [3,4]. Ac- cording to Oludumila., et al. [5], the fungus can be grown on inex- pensive substrate and is capable of producing high yield and rela- tively stable at the operating condition. Major advantage of using Citation: Adegbanke OR., et al. “Effect of pH and Temperature on the Isolation, Purification and Characterization of α-Amylase from Aspergillus niger Produced from Pigeon Pea". Acta Scientific Nutritional Health 5.12 (2021): .